α5-烟碱型乙酰胆碱受体表达下调对肺癌细胞HIF-1α表达的影响

山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (9): 8-12.

α5-烟碱型乙酰胆碱受体表达下调对肺癌细胞HIF-1α表达的影响

祖珊珊1，马晓丽1,2，郏雁飞1,2，赵云3，贾颖1，肖东杰1,2，汪运山1,2

1. 山东大学附属济南市中心医院中心实验室，济南 250013；
2. 山东省肿瘤防治靶向分子研究重点实验室，济南 250013；
3. 山东省警官总医院血液净化科，济南 250002

收稿日期:2012-12-03 出版日期:2013-09-10 发布日期:2013-09-10
通讯作者: 马晓丽，E-mail：maxL7125@yahoo.com
基金资助:
国家自然基金面上项目(81272588)；山东省自然科学基金（ZR2012HM061）；山东省高等学校科技计划（J11LC14）；国家重点基础研究发展计划（973计划）（2012CB966503-4）。

Effects of down-regulated α5-nicotinic acetylcholine receptor expression on HIF-1α expression in lung cancer cells

ZU Shan-shan1, MA Xiao-li1,2, JIA Yan-fei1,2, ZHAO Yun3, JIA Ying1, XIAO Dong-jie1,2, WANG Yun-shan1,2

1. Central Laboratory, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China；
2. Shandong Province Key Laboratory of Tumor Prevention and Target Molecule Research, Jinan 250013, China;
3. Blood Purification Division of Shandong Police General Hospital, Jinan 250002, China

Received:2012-12-03 Online:2013-09-10 Published:2013-09-10

摘要/Abstract

摘要：

目的通过下调α5-烟碱型乙酰胆碱受体（α5-nAChR）的表达，体外研究尼古丁促肺癌细胞增殖过程中α5-nAChR对缺氧诱导因子（HIF-1α）表达的影响。方法以不同浓度尼古丁作用人肺腺癌细胞株A549，分别采用实时荧光定量PCR和Western blotting检测A549细胞中α5-nAChR、HIF-1α mRNA和蛋白的表达。MTT法、实时荧光定量PCR、Western blotting检测CHRNA5-siRNA转染后，尼古丁促A549细胞增殖的作用和HIF-1α表达的变化。结果尼古丁浓度为1μmol/L时，α5-nAChR和HIF-1α表达明显升高（P<0.05），二者表达呈尼古丁浓度依赖性。CHRNA5-siRNA转染显著抑制α5-nAChR、HIF-1α表达（P<0.05），尼古丁促A549细胞增殖的作用明显受到抑制（P<0.05）。结论 α5-nAchR通过调节HIF-1α的表达，在尼古丁促肺癌细胞增殖中起作用，提示α5-nAChR/HIF-1α信号轴可能是吸烟相关性肺癌发生的重要分子机制之一。

关键词: CHRNA5基因；α5-烟碱型乙酰胆碱受体；缺氧诱导因子；尼古丁；肺肿瘤

Abstract:

Objective To explore the influence of small interfering RNA (siRNA) down-regulated α5-nicotinic acetylcholine receptor (α5-nAChR) expression on hypoxia-induced factor-1α(HIF-1α) expression during the process of nicotine-induced lung cancer A549 cell proliferation in vitro. Methods Expressions of α5-nAChR and HIF-1α at mRNA and protein levels in A549 cells were detected by FQ-PCR and Western blotting after treatment with various concentrations of nicotine. After CHRNA5-siRNA transfection, nicotine-induced A549 cell proliferation and expression of HIF-1α were detected by MTT, FQ-PCR and Western blotting respectively. Results Expressions of α5-nAChR and HIF-1α were significantly enhanced with 1μmol/L nicotine treatment compared with the other concentrations(P<0.05), which presented a nicotine dose-dependent manner. After CHRNA5-siRNA transfection, the expressions of α5-nAChR mRNA and protein decreased obviously(P<0.05). Nicotine-induced A549 cell proliferation was significantly inhibited (P＜0.05). Conclusion α5-nAchR plays an important role in nicotine-induced A549 cell proliferation via regulating expression, which suggests that α5-nAChR/HIF-1α axis may be one of the important molecular mechanisms for nicotine-related lung cancer.

Key words: CHRNA5 gene；α5-nicotinic acetylcholine receptor；Hypoxia-inducible factor-1α；Nicotine；Lung neoplasms

中图分类号:

R73-3

祖珊珊1，马晓丽1,2，郏雁飞1,2，赵云3，贾颖1，肖东杰1,2，汪运山1,2. α5-烟碱型乙酰胆碱受体表达下调对肺癌细胞HIF-1α表达的影响[J]. 山东大学学报(医学版), 2013, 51(9): 8-12.

ZU Shan-shan1, MA Xiao-li1,2, JIA Yan-fei1,2, ZHAO Yun3, JIA Ying1, XIAO Dong-jie1,2, WANG Yun-shan1,2. Effects of down-regulated α5-nicotinic acetylcholine receptor expression on HIF-1α expression in lung cancer cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(9): 8-12.

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